抗犬瘟热病毒核蛋白单克隆抗体的制备及其线性抗原表位鉴定

发布时间：2018-03-19 01:36

本文选题：犬瘟热病毒　切入点：单克隆抗体　出处：《中国预防兽医学报》2017年07期 　论文类型：期刊论文

【摘要】：为制备犬瘟热病毒(CDV)单克隆抗体(MAb),本研究用腺病毒表达的CDV截短N蛋白(aa401~aa523)免疫BALB/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行融合并筛选到2株能够稳定分泌抗CDV N蛋白的MAb,命名为N1-C8和N1-C41。经间接ELISA测定N1-C8和N1-C41培养上清液及小鼠腹水效价分别为1∶1 024、1∶106和1∶512、1∶105。通过肽扫描的方法筛选出N1-C8的抗原表位为线性表位~(440)ENQGGDKYPIHFNDE454,但并未能够筛选出N1-C41的抗原表位,推测可能是因为其抗原表位为空间构象型。Western blot结果显示N1-C8和N1-C41两株MAb在识别CDV的不同毒力病毒株上有差异,可能与CDV强弱病毒株在N蛋白的差异变化有关。本研究两株MAb的制备为CDV不同毒力株的鉴别诊断提供了可能。
[Abstract]:In order to prepare a monoclonal antibody against canine distemper virus (CDV), BALB/c mice were immunized with CDV truncated N protein (aa401a 523) expressed by adenovirus. The spleen cells of immunized mice were fused with SP2/0 myeloma cells, and two strains of MAbs which could stably secrete anti CDV N protein were selected and named N1-C8 and N1-C41.The titers of supernatants and ascites of N1-C8 and N1-C41 cultured in mice were determined by indirect ELISA, and the titers of supernatants and ascites of mice were 1: 10241: 106, respectively. The antigen epitopes of N1-C8 were obtained by peptide scanning. The epitope of N1-C8 was a linear epitope, ENQGDDKYPIHFNDE4544.However, the epitopes of N1-C41 could not be screened out. It is suggested that the antigenic epitopes of N1-C8 and N1-C41 have different virulence in different virulent strains of CDV. The results of Western blot indicate that the antigenic epitopes of N1-C8 and N1-C41 are different in different virulence virus strains. It may be related to the variation of N protein in CDV strain. The preparation of two strains of MAb provides the possibility for differential diagnosis of different virulence strains of CDV.
【作者单位】：中国农业科学院特产研究所特种经济动物分子生物学国家重点实验室;
【基金】：吉林省自然科学基金(20150101104JC)
【分类号】：S852.65

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